Karttunen T, Risteli J, Autio-Harmainen H, Risteli L
Kidney Int. 1986 Oct;30(4):586-91. doi: 10.1038/ki.1986.225.
Specific radioimmunoassays for the 7-S domain of type IV collagen and the fragment P1 of laminin were used to quantify these basement membrane proteins in human kidney cortex at different ages and in some patients with diabetes mellitus. The antigens were solubilized by treating the tissue samples with the proteolytic enzymes collagenase, trypsin and pepsin. Total collagen content (as indicated by hydroxyproline concentration) increased with age, and the proportion of the collagen that could be solubilized by any enzyme treatment decreased. The type IV collagen concentration increased significantly with age, whereas the laminin concentration tended to decrease. In the one case of a type I diabetic the amounts of both antigens exceeded those in the age matched controls. In four type II diabetics the results were comparable with those for other aged cases. The distribution of the proteins was studied using the peroxidase-antiperoxidase method. The staining intensity and thickness of both antigens increased with age in the mesangium and Bowmans capsules, the change in type IV collagen staining being more evident. In diabetic patients these changes were more pronounced and other basement membranes appeared thicker in the stainings. These results indicate that basement membrane material accumulates in the kidney cortex during aging and that an alteration takes place in the composition of the basement membranes, the proportion of type IV collagen increasing and that of laminin decreasing.
采用针对IV型胶原7-S结构域和层粘连蛋白片段P1的特异性放射免疫分析法,对不同年龄段的人肾皮质以及部分糖尿病患者肾皮质中的这些基底膜蛋白进行定量分析。通过用蛋白水解酶胶原酶、胰蛋白酶和胃蛋白酶处理组织样本,使抗原溶解。总胶原蛋白含量(以羟脯氨酸浓度表示)随年龄增长而增加,而经任何酶处理后可溶解的胶原蛋白比例下降。IV型胶原浓度随年龄显著增加,而层粘连蛋白浓度则趋于下降。在1例I型糖尿病患者中,两种抗原的含量均超过年龄匹配的对照组。在4例II型糖尿病患者中,结果与其他同龄病例相当。采用过氧化物酶-抗过氧化物酶法研究蛋白质的分布。在系膜和鲍曼囊中,两种抗原的染色强度和厚度均随年龄增加,IV型胶原染色的变化更为明显。在糖尿病患者中,这些变化更为显著,且在染色中其他基底膜显得更厚。这些结果表明,衰老过程中基底膜物质在肾皮质中积累,且基底膜的组成发生改变,IV型胶原比例增加而层粘连蛋白比例下降。